PHOTOSYNTHESIS

Cyclic photophosphorylation:
• This form of photophosphorylation occurs on the stroma lamella or fret channels. In cyclic
photophosphorylation, the high energy electron released from P700 of PS1 flow down in a cyclic pathway.
• In cyclic electron flow, the electron begins in a pigment complex called photosystem I, passes from the primary
acceptor to ferredoxin and then to plastoquinone, then to cytochrome b6f (a similar complex to that found in
mitochondria), and then to plastocyanin before returning to photosystem-I.
• This transport chain produces a proton-motive force, pumping H+ ions across the membrane; this produces a
concentration gradient that can be used to power ATP synthase during chemiomosis.
• This pathway is known as cyclic photophosphorylation, and it produces neither O2 nor NADPH. Unlike non-
cyclic photophosphorylation, NADP+ does not accept the electrons; they are instead sent back to cytochrome b6f
complex.
• Cyclic photophosphorylation occurs in both aerobic and anaerobic conditions. In this electron transport system,
the electron which was ejected from P700 molecule is cycled back, thus the process is known as cyclic electron
transport and the phosphorylation as cyclic photophosphorylation.
Non-cyclic photophosphorylation:
• Non-cyclic photophosphorylation is a two-stage process involving two different chlorophyll photosystems.
• Being a light reaction, non-cyclic photophosphorylation occurs in the thylakoid membrane.
• First, a water molecule is broken down into 2H+ 1/2 O2 + 2e− by a process called photolysis (or light-splitting). The two
electrons from the water molecule are kept in photosystem II, while the 2H+ and 1/2O2 are left out for further use. Then
a photon is absorbed by chlorophyll pigments surrounding the reaction core center of the photosystem.
• The light excites the electrons of each pigment, causing a chain reaction that eventually transfers energy to the core of
photosystem II, exciting the two electrons that are transferred to the primary electron acceptor, pheophytin. The deficit of
electrons is replenished by taking electrons from another molecule of water.
• The electrons transfer from pheophytin to plastoquinone which takes the 2e− from Pheophytin, and two H+ Ions from
the stroma and forms PQH2, which later is broken into PQ, the 2e− is released to Cytochrome b6f complex and the two
H+ ions are released into thylakoid lumen. The electrons then pass through the Cyt b6 and Cyt f. Then they are passed to
plastocyanin providing the energy for hydrogen ions (H+) to be pumped into the thylakoid space. This creates a gradient,
making H+ ions flow back into the stroma of the chloroplast, providing the energy for the regeneration of ATP.
The photosystem II complex replaced its lost electrons from an external source; however, the two other electrons
are not returned to photosystem II as they would in the analogous cyclic pathway. Instead, the still-excited
electrons are transferred to a photosystem I complex, which boosts their energy level to a higher level using a
second solar photon. The highly excited electrons are transferred to the acceptor molecule, but this time is passed
on to an enzyme called Ferredoxin-NADP+ reductase which uses them to catalyze the reaction (as shown):
NADP+ + 2H+ + 2e− → NADPH + H+
This consumes the H+ ions produced by the splitting of water, leading to a net production of molecular O2, ATP,
and NADPH+H+ with the consumption of solar photons and water. The concentration of NADPH in the
chloroplast may help regulate which pathway electrons take through the light reactions. When the chloroplast runs
low on ATP for the Calvin Cycle, NADPH will accumulate and the plant may shift from noncyclic to cyclic
electron flow.
This pathway is also called as the ‘Z-scheme of photosynthesis’ because the redox diagram from
P680 to P700 looks like a big alphabet ‘Z’. In this scheme of electron transport, the electron
ejected from PSII did not return to its place of origin, instead it was passed on to PSI. Similarly,
the electron emitted from PSI did not cycle back but was used to reduce NADP+ into NADPH +
H+. Therefore, this electron transport has been called as Non-Cyclic electron transport and the
phosphorylation as Non- Cyclic Photophosphorylation. Thus in this system, the PSI is reduced by
electrons coming from PSII and the PSII is reduced by electrons coming from water. This is
associated with photo-oxidation of water releasing molecular O2.
Significance of Non-Cyclic Photophosphorylation
Non-Cyclic electron transport/ Photophosphorylation comprises of the sequence of electron
transport where NADP+ is reduced by PSI, the PSI is reduced by PSII and finally PSII is
reduced by electrons coming from photo-oxidation of water. Thus the cycle is broken during
this electro transport and therefore, it is called non-cyclic. The photo-oxidation of water and
evolution of molecular oxygen is associated with this type of electron transport. It released a
major portion of protons which finally generate proton motive force to produce ATP. The
non- cyclic electron transport is most important in photosynthesis as it supplies assimilatory
power in the form of NADPH and ATP for CO2 assimilation and purifies the atmospheric
air.
Chemiosmotic Hypothesis
The chemiosmotic hypothesis has been put forward to explain the mechanism. Like
in respiration, in photosynthesis too, ATP synthesis is linked to development of a
proton gradient across a membrane. This time these are the membranes of
thylakoid. There is one difference though, here the proton accumulation is towards
the inside of the membrane, i.e., in the lumen. In respiration, protons accumulate in
the intermembrane space of the mitochondria when electrons move through the ETS.
Let us understand what causes the proton gradient across the membrane. We need
to consider again the processes that take place during the activation of electrons and
their transport to determine the steps that cause a proton gradient to develop (Figure
13.7).
(a) Since splitting of the water molecule takes place on the inner side of the
membrane, the protons or hydrogen ions that are produced by the splitting of water
(b) As electrons move through the photosystems, protons are transported across the
membrane. This happens because the primary accepter of electron which is located
towards the outer side of the membrane transfers its electron not to an electron carrier but
to an H carrier. Hence, this molecule removes a proton from the stroma while transporting
an electron. When this molecule passes on its electron to the electron carrier on the inner
side of the membrane, the proton is released into the inner side or the lumen side of the
membrane.
(c) The NADP reductase enzyme is located on the stroma side of the membrane. Along
with electrons that come from the acceptor of electrons of PS I, protons are necessary for
the reduction of NADP+ to NADPH+ H+. These protons are also removed from the stroma.
Hence, within the chloroplast, protons in the stroma decrease in number, while in the
lumen there is accumulation of protons. This creates a proton gradient across the thylakoid
membrane as well as a measurable decrease in pH in the lumen.
This gradient is important because it is the breakdown of this gradient that leads to the
synthesis of ATP. The gradient is broken down due to the movement of protons across the
membrane to the stroma through the trans membrane channel of the CF0 of the ATP
synthase. The ATP synthase enzyme consists of two parts: one called the CF0 is
embedded in the thylakoid membrane and forms a transmembrane channel that carries
out facilitated diffusion of protons across the membrane. The other portion is called CF1
and protrudes on the outer surface of the thylakoid membrane on the side that faces the
stroma. The break down of the gradient provides enough energy to cause a
conformational change in the CF1 particle of the ATP synthase, which makes the enzyme
synthesise several molecules of energy packed ATP.
Chemiosmosis requires a membrane, a proton pump, a proton gradient and ATP
synthase. Energy is used to pump protons across a membrane, to create a
gradient or a high concentration of protons within the thylakoid lumen. ATP
synthase has a channel that allows diffusion of protons back across the
membrane; this releases enough energy to activate catalyses the formation of
ATP. Along with the NADPH produced by the movement of electrons, the ATP will
be used immediately in the biosynthetic reaction taking place in the stroma,
responsible for fixing CO2, and synthesis of sugars.
Calvin cycle or C3 cycle
It is a cyclic reaction occurring in the dark phase of photosynthesis. In
this reaction, CO2 is converted into sugars and hence it is a process of
carbon fixation. The Calvin cycle was first observed by Melvin Calvin,
Benson and Basham in chlorella, unicellular green algae. Calvin was
awarded Nobel Prize for this work in 1961. Since the first stable
compound in Calvin cycle is a 3 carbon compound (3 phosphoglyceric
acid), the cycle is also called as C3 cycle. The reactions of Calvin’s
cycle occur in three phases.
1. Carboxylative phase
2. Reductive phase
The CO2 fixation pathway has several
names:
•The reductive pentose phosphate (RPP)
pathway.
•The C3 pathway.
•The photosynthetic carbon reduction
cycle.
•The Calvin cycle
Stage 1, Carboxylative Phase: Ribulose 1,5-Bisphosphate Carboxylase-Oxygenase (Rubisco)
• Gaseous CO2 and the 5-carbon sugar ribulose 1,5- bisphosphate forms 3-phosphoglycerate
• Reaction is metabolically irreversible (G0’ = -51.9 kJ/mole)
• Rubisco makes up about 50% of the soluble protein in plant leaves, and is one of the most
abundant enzymes in nature and is present on the outer surface of thylakoid membrane
Six molecules of CO2 are accepted by 6 molecules of 5C compound viz., ribulose 1,5 biphosphate to
form six molecules of an unstable intermediate 6C compound. This reaction is catalyzed by the
enzyme, Ribulose 1,5-Bisphosphate Carboxylase-Oxygenase (Rubisco)/carboxy dismutase. The six
molecules of the unstable 6 carbon compound are converted by the addition of 6 molecules of water
into 12 molecules of 3 phosphoglyceric acid (3C compound). This reaction is also catalyzed by the
enzyme Rubisco/carboxy mutase. 3 phosphoglyceric acid (PGA) is the first stable product of dark
reaction of photosynthesis and since it is a 3 carbon compound, this cycle is known as C3 cycle .
2. Reductive phase
12 molecules of 3-PGA are phosphorylated by 12 molecules of ATP (produced in the light
reaction) to yield 6 molecules of 1,3 diphosphoglyceric acid and 12 molecules of ADP. This
reaction is catalyzed by the enzyme, Phosphoglycerate Kinase

12 molecules of 1, 3 diphosphoglyceric acid are reduced with the use of 12 molecules of

NADPH + H+ (produced in light reaction) to form 12 molecules of 3 phosphoglyceraldehyde
(PGAL) or Glyceraldehyde-3-phosphate. This reaction is catalysed by the enzyme, triose
phosphate dehydrogenase.
3. Regenerative phase
In the regenerative phase, the ribose bisphosphate is regenerated. The
regenerative phase is called as pentose phosphate pathway or hexose
monophophate shunt. It involves the following steps.
1. Some of the molecules (5 mol.) out of 12 moles of 3 phosphoglyceraldehyde
isomerised into 5 moles of dihydroxy acetone phosphate in presence of Triose
phosphate isomerase. 3 moles of phosphoglyceraldehyde and 3 moles of
dihydroxyacetone phosphate then unite in the presence of the enzyme,
aldolase to form 3 moles of fructose, 1-6 bisphosphate.
2. 3 moles of Fructose 1, 6 bisphosphate is converted into 3 moles of
fructose 6 phosphate in the presence of Fructose 1,6 bisphosphatase.
Phophatase

3. 4 out of 7 moles of 3 phospho glyceraldehyde instead of forming hexose

sugars are diverted to regenerate ribulose 1-5 diphosphate
4. 2 moles of 3 phospho glyceraldehyde reacts with 2 moles of fructose 6
phosphate in the presence of enzyme transketolase to form 2 moles of
erythrose 4 phosphate ( 4C sugar) and 2 moles of xylulose 5 phosphate(5C
sugar).
5. 2 moles of Erythrose 4 phosphate combines with 2 moles of dihydroxy
acetone phosphate in the presence of the enzyme transaldolase to form
sedoheptulose 1,7 bisphosphate(7C sugar).

6. 2 moles of Sedoheptulose 1, 7 bisphosphate loses one phosphate

group in the presence of the enzyme sedoheptulose 1,7 bisphosphatase
to form 2 moles of sedoheptulose 7 phosphate.
7. 2 moles of Sedoheptulose 7-phosphate reacts with 2 moles of 3 phospho
glyceraldehyde in the presence of transketolase to form 2 moles of xylulose
5 phosphate and 2 moles of ribose 5 phosphate.

8. 2 moles of Ribose 5 phosphate is converted into 2 moles of ribulose 5-

phosphate in the presence of enzyme, phosphopentose isomerase. 4
molecules of xylulose phosphate are epimerised into four molecules of
ribulose 5-phosphate by Phosphopentose epimerase. Hence a total of 6
moles of Ribulose-5-phosphate is formed
9. 6 moles of Ribulose 5 phosphate is converted into 6 moles of ribulose
1, 5 bisphosphate in the presence of enzyme, phosphoribulose kinase
and 6 ATP molecules. Thus RuBP is regenerated and can combine with
CO2 to continue the cycle.
C4 cycle or Hatch and Slack pathway
It is the alternate pathway of C3 cycle to fix CO2. In this cycle, the first formed stable
compound is a 4 carbon compound viz., oxaloacetic acid. Hence it is called C4
cycle. The pathway is also called as Hatch and Slack as they worked out the pathway
in 1966 and it is also called as C4 dicarboxylic acid pathway. This pathway is
commonly seen in many grasses, sugar cane, maize, sorghum and amaranthus.
The C4 plants show a different type of leaf anatomy. The chloroplasts are dimorphic
in nature. In the leaves of these plants, the vascular bundles are surrounded by
bundle sheath of larger parenchymatous cells. These bundle sheath cells have
chloroplasts. These chloroplasts of bundle sheath are larger, lack grana and contain
starch grains. The chloroplasts in mesophyll cells are smaller and always contain
grana. This peculiar anatomy of leaves of C4 plants is called Kranz anatomy. The
bundle sheath cells are bigger and look like a ring or wreath. Kranz in German means
The C4 cycle involves two carboxylation reactions, one taking place in
chloroplasts of mesophyll cells and another in chloroplasts of bundle sheath
cells. There are four steps in Hatch and Slack cycle:
1. Carboxylation
2. Breakdown
3. Splitting
4. Phosphorylation
1. Carboxylation
It takes place in the chloroplasts of mesophyll cells. Phosphoenolpyruvate, a 3 carbon compound
picks up CO2 and changes into 4 carbon oxaloacetate in the presence of water. This reaction is
catalysed by the enzyme, phosphoenol pyruvate carboxylase (PEPCo).

2. Breakdown
Oxaloacetate breaks down readily into 4 carbon malate and aspartate in the presence of the enzyme,
transaminase and malate dehydrogenase. These compounds diffuse from the mesophyll cells into
sheath cells.
3. Splitting
In the sheath cells, malate and aspartate split enzymatically to yield free
CO2 and 3 carbon pyruvate. The CO2 is used in Calvin’s cycle in the sheath
cell.

The second Carboxylation occurs in the chloroplast of bundle sheath cells.

The CO2 is accepted by 5 carbon compound ribulose 1,5- bisphosphate in
the presence of the enzyme, Rubisco and ultimately yields 3
phosphoglyceric acid. Some of the 3 phosphoglyceric acid is utilized in the
4. Phosphorylation
The pyruvate molecule is transferred to chloroplasts of mesophyll cells where,
it is phosphorylated to regenerate phosphoenol pyruvate in the presence of
ATP. This reaction is catalysed by pyruvate phosphokinase and the
phophoenol pyruvate is regenerated.

In Hatch and Slack pathway, the C3 and C4 cycles of carboxylation are linked
and this is due to the Kranz anatomy of the leaves. The C4 plants are more
efficient in photosynthesis than the C3 plants. The enzyme, phosphoenol
pyruvate carboxylase of the C4 cycle is found to have more affinity for CO2
than the ribulose diphosphate carboxylase of the C3 cycle in fixing the
molecular CO2 in organic compound during Carboxylation.
Crassulacean Acid Metabolism (CAM) cycle or the dark fixation of CO 2 in succulents

CAM is a cyclic reaction occurring in the dark phase of photosynthesis in the plants of Crassulaceae. It

is a CO2 fixation process wherein, the first product is malic acid. It is the third alternate pathway of

Calvin cycle, occurring in mesophyll cells. The plants exhibiting CAM cycle are called CAM plants. Most

of the CAM plants are succulents e.g., Bryophyllum, Kalanchoe, Crassula, Sedium, Kleinia etc. It is

also seen in certain plants of Cactus e.g. Opuntia, Orchid and Pineapple families.

CAM plants are usually succulents and they grow under extremely xeric conditions. In these plants,

the leaves are succulent or fleshy. The mesophyll cells have larger number of chloroplasts and the

vascular bundles are not surrounded by well defined bundle sheath cells.

In these plants, the stomata (Scotoactive) remain open during night and closed during day time. The

CAM plants are adapted to photosynthesis and survival under adverse xeric conditions. CAM plants

are not as efficient as C4 plants in photosynthesis. But they are better suited to conditions of extreme
CAM involves two steps:
1. Acidification
2. Deacidification
Acidification
In darkness, the stored carbohydrates are converted into phophoenol pyruvic acid. The stomata in
CAM plants are open in dark and they allow free diffusion of CO 2 from the atmosphere into the leaf.
Now, the phosphoenolpyruvic acid carboxylated by the enzyme phosphoenol pyruvic acid carboxylase
and is converted in to oxalaoacetic acid.

The oxaloacetic acid is then reduced to malic acid in the presence of the enzyme malate
dehydrogenase. The reaction requires NADPH+ H+.

The malic acid produced in dark is stored in the vacuole. The malic acid increases the acidity of the
Deacidification
During day time, when the stomata are closed, the malic acid is
decarboxylated to produce pyruvic acid and evolve carbon dioxide in the
presence of the malic enzyme. When the malic acid is removed, the acidity
decreases in the cells. This is called deacidification. One molecule of NADP+
is reduced in this reaction.

The pyruvic acid may be oxidized to CO2 by the pathway of Kreb’s cycle or it
may be reconverted to phosphoenol pyruvic acid and synthesize sugar by C3
cycle. The CO2 released by deacidification of malic acid is accepted by
CAM is a most significant pathway in succulent plants. The stomata are closed
during day time to avoid transpiration loss of water. As the stomata are closed,
CO2 cannot enter into the leaves from the atmosphere. However, they can carry
out photosynthesis during the day time with the help of CO2 released from
organic acids. During night time, organic acids are synthesized in plenty with the
help of CO2 released in respiration and the CO2 entering from the atmosphere
through the open stomata. Thus, the CO2 in dark acts as survival value to these
plants.
During the CAM cycle, Since stomata is only opened during night in CAM plants,
CO2 can enter the mesophyll cells and its acceptor, PEP is carboxylated to
oxaloacetic acid and finally reduced to malic acid and stored in vacuole during
night and this malic acid is only used during day time when light reactions start
producing ATP and assimilatory power, NADPH+H+, until then the CAM cycle
PHOTORESPIRATION
The excessive respiration that takes place in green cells in the presence of light is called
as photorespiration. Decker (1955) discovered the process and it is also called as C2
cycle as the 2 carbon compound glycolic acid acts as the substrate in photorespiration.
In general, respiration takes place under both light and dark conditions. However in
some plants, the respiration is more in light than in dark. It is 3-5 times higher than the
rate of respiration in dark. Photorespiration is carried out only in the presence of light.
But the normal respiration is not light dependent and it is called dark respiration.
In photorespiration, temperature and oxygen concentration play an important role.
Photorespiration is very high when the temperature is between 25 and 30 ºC. The rate of
photorespiration increases with the increase in the concentration of oxygen. Three cell
organelles namely chloroplast, peroxisome and mitochondria are involved in the
photorespiration. This kind of respiration is seen in plants like cotton, pulses, capsicum,
peas, tomato, petunia, soybean, wheat, oats, paddy, chlorella etc and it is absent in
Mechanism
1. In the presence of excess oxygen and low CO2 , ribulose 1,5 diphosphate produced in the
chloroplast during photosynthesis is split into 2 phospho glycolic acid and 3 phosphoglyceric
acid by the enzyme, ribulose 1,5 diphosphate oxygenase
2. The 3 phospho glyceric acid enters the Calvin cycle.
3. In the next step, phosphate group is removed from 2 phosphoglycolic acid to produce
glycolic acid by the enzyme, 2-phosphoglycolate phosphatase.
4. Glycolic acid then it come out of chloroplast and enter the peroxisome. Here, it combines
with oxygen to form glyoxylic acid and hydrogen peroxide. This reaction is catalyzed by the
enzyme, glycolic acid oxidase. Hydrogen peroxide is toxic and it is broken down into water
and oxygen by the enzyme, Catalase. Photorespiration is an oxidation process. In this
process, glycolic acid is converted into carbohydrate and CO2 is released as the by product.
As glycolic acid is oxidized in photorespiration, it is also called as glycolate metabolism.
5. The glyoxylic acid converted into glycine by the addition of one amino group with the help
of the enzyme, glutamate-glyoxylate amino transferase.
6. Now, the glycine is transported from the peroxisome into the mitochondria. In the
mitochondria, two molecules of glycine condense to form serine and liberate carbon
dioxide and ammonia in presence of glycine decarboxylase complex.
7. Amino group is removed from serine to form hydroxy pyruvic acid in the presence
of the enzyme, serine glyoxylate transaminase.
8. Hydroxy pyruvic acid undergoes reduction with the help of NADH to form glyceric
acid in the presence of enzyme alpha hydroxy pyruvate reductase.
9. Finally, regeneration of 3 phosphoglyceric acid occurs by the phosphorylation of
glyceric acid with ATP. This reaction is catalyzed by the enzyme, glycerate kinase.
10. The 3 phosphoglyceric acid is an intermediate product of Calvin cycle. If it enters
the chloroplast, it is converted into carbohydrate by photosynthesis and it is
suppressed nowadays with the increased CO2 content in the atmosphere.